Abstract
Colony-forming assays represent prospective methods, where cells isolated from enzymatically dissociated tissues or from tissue cultures are assessed for their proliferative capacity in vitro. Complex tissues such as the epithelial component of the skin (the epidermis) are characterized by a substantial cellular heterogeneity. Analysis of bulk populations of cells by colony-forming assays can consequently be convoluted by a number of factors that are not controlled for in population wide studies. It is therefore advantageous to refine in vitro growth assays by sub-fractionation of cells using flow cytometry. Using markers that define the spatial origin of epidermal cells, it is possible to interrogate the specific characteristics of subpopulations of cells based on their in vivo credentials. Here, we describe how to isolate, culture, and characterize keratinocytes from murine back and tail skin sorted by surface antigens associated with adult stem cell characteristics.
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Acknowledgment
This work was supported by The Danish Cancer Society, The Lundbeck Foundation, The Novo Nordic Foundation, Leo Pharma Foundation, and The A.P. Møller Foundation for the Advancement of Medical Science. K.B.J. is an EMBO young investigator. We thank past and present members of the Jensen lab for input, ideas, and contributions to the optimized protocols.
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Moestrup, K.S., Andersen, M.S., Jensen, K.B. (2017). Isolation and In Vitro Characterization of Epidermal Stem Cells. In: Di Nardo, P., Dhingra, S., Singla, D. (eds) Adult Stem Cells. Methods in Molecular Biology, vol 1553. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6756-8_6
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DOI: https://doi.org/10.1007/978-1-4939-6756-8_6
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